Circular miRNA Sponges with Bulged Binding Sites
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Solution Overview
Problem
Current miRNA interference technologies face challenges such as short half-lives, off-target effects, and potential accumulation of non-metabolizable molecules, necessitating improved methods for therapeutic applications, particularly in treating diseases like heart failure where miR-212/132 family plays a significant role in cardiac hypertrophy.
Innovation Solution
The development of engineered circular miRNA sponges, termed circmiRs, with optimized binding sites and spacers, designed to specifically target miR-212/132, offering enhanced stability and efficacy compared to linear counterparts, and demonstrating cardiomyocyte-specific delivery in vivo.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If linear miRNA sponge constructs are used, then miRNA binding function is achieved, but the constructs are susceptible to exonucleolytic degradation resulting in short half-lives
Solution Approach 1:
The patent applies circularization of the miRNA sponge construct, transforming the linear RNA molecule into a closed circular structure. This curvature eliminates free ends that are targets for exonucleases, thereby conferring resistance to exonucleolytic degradation and extending the half-life of the sponge construct in biological systems
2Adaptability or versatility
If multiple binding sites for different miRNAs are incorporated, then functional class inhibition is achieved, but the construct complexity increases
Solution Approach 1:
The circular miRNA sponge construct is designed with multiple distinct binding sites that can simultaneously accommodate different miRNA sequences. This multi-functional design allows a single construct to inhibit multiple miRNA types and their associated functional classes, thereby achieving versatility without requiring separate constructs for each target
Solution Approach 2:
The sponge construct is segmented into multiple modular binding sites separated by non-identical spacers. Each binding site functions as an independent module that can be optimized for specific miRNA targets, while the overall circular structure integrates these segments into a cohesive multi-functional platform
3Measurement precision
If conventional miRNA interference technology is used, then miRNA targeting is achieved, but off-target effects and accumulation of non-metabolisable molecules occur
Solution Approach 1:
The patent employs chemical modification parameters of the nucleotide building blocks in the circular sponge construct. By altering chemical parameters such as sugar ring modifications or phosphate backbone changes, the construct achieves enhanced metabolic degradability while maintaining specific binding affinity for target miRNAs, thereby reducing off-target effects and toxic accumulation
Data Source
AI summary
The present invention relates to miRNA interference technology. More specifically the invention relates to circular miRNA sponges that carry a plurality of binding sites directed to at least two types of miRNA and separated by random, non-identical spacers, allowing for the inhibition of functional classes of m1RNAs. Preferably, the binding sites are bulged binding sites wherein each bulge is created by a one base deletion and two base mismatch at positions 9-11 nt from the 3′ end of each binding site. Preferably, each spacer is 6 to 24 nucleotides in length. Preferably, the binding sites are against miR-132 and miR-212, miR-17-5p and miR-18a-5p, or miR-20b-5p and miR-106a-5p. Construction vectors and uses of said miRNA sponges for the treatment of diseases, such as cardiomyopathy and cancer, are also disclosed.